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目的:研究胶原诱导的关节炎(CIA)大鼠关节中局部浸润的T细胞TCRVβ克隆型。方法:用II型胶原(CII)诱导Lewis大鼠发生关节炎后,经流式细胞术分析大鼠外周血CD4+和CD8+细胞的变化。提取CIA大鼠关节组织的总RNA,通过RT-PCR/SSCP方法分析大鼠两后肢足关节中TCRVβ克隆型的一致率。将CIA大鼠脾淋巴细胞转移至同系正常大鼠,观察发病情况。结果:从注射CII的第30天起,大鼠后爪出现红、肿等症状。流式细胞术分析显示,与正常大鼠相比较,CIA大鼠外周血CD8+T细胞的降低有显著性意义(P<0.01),CD4+T/CD8+T的比值大于2(P<0.01),提示CIA大鼠体内存在T淋巴细胞亚群的比例失调。CIA大鼠两后肢足关节中聚集的TCRVβ克隆型的一致率,以TCRVβ5.2和8.2最高,分别为78.89%和72.23%。脾淋巴细胞转移试验显示,受者发病的大鼠关节中,出现与供者CIA大鼠一致的TCRVβ克隆型条带,表明关节局部聚集的TCRVβ克隆型与关节炎(RA)的发病有关。结论:CIA大鼠关节局部存在以TCRVβ5.2和8.2为主的致病性的T细胞TCRVβ克隆型,可将其作为治疗CIA的靶物质,进行实验动物RA治疗的研究。
AIM: To investigate the TCRVβ clonotype of locally infiltrating T cells in the joints of collagen-induced arthritis (CIA) rats. Methods: After induced by arthritis in Lewis rats with type II collagen (CII), the changes of CD4 + and CD8 + cells in peripheral blood of rats were analyzed by flow cytometry. Total RNA was extracted from the joint tissues of CIA rats and the concordance rates of TCRVβ clonotypes in the two hind paws of the rats were analyzed by RT-PCR / SSCP. The splenic lymphocytes of CIA rats were transferred to normal rats, observed the incidence. Results: From the 30th day after CII injection, the hindpaws of the rats appeared red, swollen and other symptoms. Flow cytometry analysis showed that compared with normal rats, the decrease of CD8 + T cells in CIA rats was significant (P <0.01), and the ratio of CD4 + T / CD8 + T was greater than 2 (P <0.01) ), Suggesting that there is an imbalance of T lymphocyte subsets in CIA rats. The concordance rates of TCRVβ clonality clustered in the two hind paws of CIA rats were the highest with TCRVβ5.2 and 8.2, which were 78.89% and 72.23% respectively. Splenic lymphocyte metastasis tests showed that clonal TCRVβ clones consistent with donor CIA rats appeared in the affected rat joints, indicating that the locally aggregated TCRVβ clonoid in the joints is associated with the onset of arthritis (RA). CONCLUSION: TCRVβ clonotype of TCRVβ5.2 and 8.2, which are mainly TCRVβ5.2 and 8.2, exists locally in the joints of CIA rats and can be used as a target substance for the treatment of CIA to study the treatment of RA in experimental animals.